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Abstract:

An automated method for analyzing seeds generally includes collecting
image data from individual seeds using a seed sampling system,
determining at least one characteristic of each of the individual seeds
based on the collected image data, and removing tissue from each of the
individual seeds using the seed sampling system. The method also
includes, prior to removing the tissue sample from each of the individual
seeds, adjusting at least one operational parameter of the seed sampling
system based on the at least one characteristic of the seed from which
the tissue is to be removed to thereby allow for generally consistent
removal of tissue from each of the individual seeds. In some aspects, the
method further includes analyzing the tissue removed from the seeds for
presence or absence of at least one characteristic, and selecting seeds
based on presence or absence of the at least one characteristic.

Claims:

1. An automated method for removing tissue from seeds using a seed
sampling system, the method comprising: collecting image data for a seed
isolated from a bulk of seeds; determining at least one characteristic of
the seed based on the collected image data; adjusting operation of the
seed sampling system based on the at least one characteristic of the
seed; and removing tissue from the seed; whereby adjusting operation of
the seed sampling system helps provide generally consistent removal of
tissue from multiple different seeds in the bulk of seeds.

2. The method of claim 1, further comprising isolating the seed from the
bulk of seeds.

3. The method of claim 2, further comprising directing the isolated seed
to one of multiple seed holders of a seed transport.

4. The method of claim 1, further comprising: receiving the tissue
removed from the seed in a sample tray and mapping a location of the
tissue in the sample tray; and/or receiving the seed from which the
tissue is removed in a seed tray and mapping a location of the seed in
the seed tray.

5. The method of claim 1, further comprising collecting the tissue
removed from the seed and correlating the collected tissue to the seed.

6. The method of claim 1, further comprising screening the tissue removed
from the seed for presence or absence of at least one characteristic.

7. The method of claim 1, wherein each of collecting image data for a
seed, determining at least one characteristic of the seed based on the
collected image data, adjusting operation of the seed sampling system
based on the at least one characteristic of the seed, and removing tissue
from the seed comprises automated operations.

8. The method of claim 1, wherein the at least one characteristic
includes size of the seed; and wherein adjusting operation of the seed
sampling system includes adjusting operation of the seed sampling system
based on the size of the seed so that a predetermined amount of tissue is
removed from the seed; whereby the seed sampling system is thereby
capable of removing generally equal amounts of tissue from seeds in the
bulk of seeds having different sizes.

9. A high-throughput, automated method for use in analyzing seeds, the
method comprising: collecting image data from multiple individual seeds
using a seed sampling system; determining at least one characteristic of
each of the individual seeds based on the collected image data; removing
tissue from each of the individual seeds using the seed sampling system;
and prior to removing the tissue sample from each of the individual
seeds, adjusting at least one operational parameter of the seed sampling
system based on the at least one characteristic of the seed from which
the tissue is to be removed to thereby allow for generally consistent
removal of tissue from each of the multiple individual seeds.

10. The method of claim 9, further comprising isolating the multiple
individual seeds from a bulk of seeds.

11. The method of claim 10, further comprising directing each of the
isolated seeds to one of multiple seed holders of a seed transport.

12. The method of claim 9, further comprising: receiving the tissue
removed from the seeds in a sample tray and mapping locations of the
tissue in the sample tray; and/or receiving the seeds from which the
tissue is removed in a seed tray and mapping locations of the seeds in
the seed tray.

13. The method of claim 9, further comprising collecting the tissue
removed from the seeds and correlating the collected tissue to the seeds
from which it was removed.

14. The method of claim 9, further comprising analyzing the tissue
removed from the seeds for presence or absence of at least one
characteristic.

15. The method of claim 14, further comprising selecting desired ones of
the individual seeds based on presence or absence of the at least one
characteristic.

16. The method of claim 9, wherein the operations of collecting image
data, determining at least one characteristic, removing tissue, and
adjusting at least one operational parameter of the seed sampling system
comprise automated operations.

17. The method of claim 9, wherein the at least one characteristic of
each of the individual seeds includes size of the seeds; and wherein
adjusting at least one operational parameter of the seed sampling system
includes adjusting operation of a cutting device of the seed sampling
system based on the size of the seed from which the tissue is to be
removed so that generally equal amounts of tissue are removed from each
of the multiple individual seeds.

18. The method of claim 17, wherein adjusting operation of a cutting
device of the seed sampling system based on the size of the seed from
which the tissue is to be removed includes adjusting an angular rotation
of the cutting device based on the size of the seed from which the tissue
is to be removed.

19. A method for monitoring operation of an automated seed sampling
system having a seed loading station, a seed transport subsystem, and a
seed sampling station, the method comprising: sensing if individual seeds
are successfully isolated from a bulk of seeds at the seed loading
station; and sensing if the isolated seeds are properly positioned by the
seed transport subsystem adjacent the seed sampling station in
preparation for removing tissue from the isolated seeds.

20. The method of claim 19, wherein the seed sampling system further
includes a seed imaging station and a seed orientation station, the
method further comprising: sensing if the isolated seeds are properly
positioned by the seed transport subsystem adjacent the seed imaging
station in preparation for collecting image data of the isolated seeds;
sensing if the isolated seeds are properly positioned by the seed
transport subsystem adjacent the seed orientation station in preparation
for orienting the isolated seeds in a desired orientation; and sensing if
the isolated seeds are properly oriented at the seed orientation station.

[0002] This disclosure generally relates to systems and methods for taking
samples from biological materials such as seeds.

BACKGROUND

[0003] The statements in this section merely provide background
information related to the present disclosure and may not constitute
prior art.

[0004] In plant development and improvement, genetic improvements are made
in the plant, either through selective breeding or genetic manipulation,
and when a desirable improvement is achieved, a commercial quantity is
developed by planting and harvesting seeds over several generations. Not
all seeds express the desired traits and, thus, these seeds need to be
culled from the population. To hasten the process of bulking up the
population, statistical samples are taken and tested to cull seeds from
the population that do not adequately express the desired trait.

[0006] The present disclosure relates to systems and methods of separating
seeds from a plurality of seeds, extracting a sample from each seed,
sorting the extracted samples and corresponding seeds respectively to
wells in sample trays and seed trays, and mapping the respective wells to
track each sample with the seed from which it was extracted. The methods
are particularly adapted for automation, which permits a greater sampling
and sorting efficiency and throughput rate than was previously practical.

[0007] In various embodiments, the present disclosure provides an
automated system for sampling and sorting at least one seed from a
plurality of seeds. The system includes a seed loading station for
separating at least one seed from a plurality of seeds in a bulk seed
hopper, an imaging station for collecting image data of the at least one
seed, and a seed orientation station for independently positioning and
retaining each seed in a desired orientation based on the collected image
data. The system also includes a seed sample and sort station for
extracting a tissue sample from each seed, sorting each tissue sample to
a sample tray and sorting each sampled seed to a seed tray.

[0008] In various other embodiments, the present disclosure provides an
automated, high-throughput method for extracting sample material for
testing from individual seeds in a population of seeds. The method
includes separating at least one seed from a plurality of seeds in a
population, collecting image data fro the at least one seed,
independently positioning the at least one seed in a desired orientation
based on the collected image data, extracting a tissue sample from the at
least one seed, and sorting the tissue sample to a sample tray and
sorting the sampled seed to a seed tray.

[0009] In yet other various embodiments, the present disclosure provides
an automated system for sequentially removing sample material from
individual ones of a plurality of seeds while preserving the germination
viability of the seeds. The system includes a seed loading station for
separating and retaining sets of seeds from a plurality of seeds in a
bulk seed hopper, an imaging station for collecting image data of the
retained sets of seeds, and a seed orientation station for independently
positioning each seed in each seed set in a desired orientation based on
the collected image data. The system also includes a seed sample and sort
station for extracting a tissue sample from each seed in each seed set,
sorting each tissue sample to a sample tray and sorting each sampled seed
to a seed tray.

[0010] In still yet other various embodiments, the present disclosure
provides an automated, high-throughput method for sequentially extracting
sample material for testing from a plurality of seeds while preserving
the germination viability of the seeds. The method includes separating
sets of seeds from a plurality of seeds in a bulk seed hopper. Each set
of seeds is then presented for retention by a respective one of a
plurality of rotary vacuum cup banks at a seed loading station of a seed
sampling and sorting system. Each rotary vacuum cup bank includes a
plurality of rotary vacuum cup devices. The method additionally includes
collecting image data of each set of seeds retained by each rotary vacuum
cup bank at an imaging station of the seed sampling and sorting system.
The method further includes independently positioning each seed in the
set in a desired orientation based on the collected image data at a seed
orientation station of the seed sampling and sorting system. The method
still further includes extracting a sample from each seed in each set of
seeds; and sorting each sample to a sample tray and sorting each sampled
seed to a seed tray at a seed sample and sort station of the seed
sampling and sorting system.

[0011] In still other various embodiments, the present disclosure provides
a seed loading station of an automated seed sampling and sorting system.
The seed loading station includes a separating wheel for separating seeds
from a plurality of seeds in a bulk hopper. Additionally, the seed
loading station includes a tube shuttle having a plurality of first
transfer tubes extending from a plurality of openings in the tube
shuttle. The tube shuttle is structured and operable to incrementally
positioning each of the first transfer tubes under the separating wheel
such that each of the first transfer tubes receives a seed from the
separating wheel.

[0012] In other various embodiments, the present disclosure provides a
seed sample and sort station of an automated seed sampling and sorting
system. The seed sample and sort station includes a press plate bank
including a number of press plates equal to a number of seed retention
devices of an automated seed sampling and sorting system. Each seed
retention device retains a respective seed. The seed sample and sort
station additionally includes a linear actuator to which the press plate
bank is mounted. The linear actuator is controllable to lower the press
plate bank such that each press plate engages a friction plate of a
corresponding one of the retention devices and moves the respective seeds
downward to a sampling location. The seed sample and sort station further
includes a plurality of independently controlled grip and chip
assemblies. Each grip and chip assembly includes a seed gripping
mechanism for firmly holding a respective one of the seeds at the
respective sampling location as a sample is extracted from the respective
seed. Each grip and chip assembly additionally includes a sample
extraction mechanism for extracting the sample from each respective seed.

[0013] In still other embodiments, methods are provided for removing
tissue from multiple individual seeds. In one example embodiment, a
method for removing tissue from multiple individual seeds generally
includes loading multiple individual seeds in a seed transport, orienting
the multiple individual seeds in the seed transport substantially
simultaneously, and removing tissue from the oriented multiple individual
seeds.

[0014] In another example embodiment, an automated method for sampling
seeds includes separating individual seeds from a plurality of seeds,
imaging the separated seeds, and removing tissue samples from the imaged
seeds.

[0015] In other embodiments, seed sampling systems are provided. In one
example embodiment, a seed sampling system generally includes a seed
transport configured to hold multiple individual seeds together as a
group and transport the multiple individual seeds together as the group,
and to allow the multiple individual seeds to be oriented while the
multiple individual seeds are being held in the seed transport; and a
seed sampling subsystem configured to remove tissue from the oriented
multiple individual seeds.

[0016] In another example embodiment, an automated system for sampling
seeds includes a seed loading station for separating individual seeds
from a plurality of seeds held in a seed hopper, an imaging station
configured to receive the separated seeds from the seed loading station
and collect image data of the received seeds, and a seed sampling
subsystem configured to remove tissue samples from the seeds after the
image data of the seeds is collected.

[0017] Further areas of applicability of the present teachings will become
apparent from the description provided herein. It should be understood
that the description and specific examples are intended for purposes of
illustration only and are not intended to limit the scope of the present
teachings.

DRAWINGS

[0018] The drawings described herein are for illustration purposes only
and are not intended to limit the scope of the present teachings in any
way.

[0019] FIG. 1 is an isometric view of a seed sampling system in accordance
with various embodiments of the present disclosure.

[0020]FIG. 2 is a block diagram of a top view of the seed sampling system
shown in FIG. 1.

[0021]FIG. 3A is an isometric view of a seed loading station (absent
system support structure) of the seed sorter system shown in FIG. 1, in
accordance with various embodiments of the present disclosure.

[0022]FIG. 3B is a schematic of a queuing stack of the seed loading
station shown in FIG. 3A, in accordance with various embodiments of the
present disclosure.

[0023] FIG. 3C is a sectional side view of an elevator hopper of the seed
loading station shown in FIG. 3A, in accordance with various embodiments
of the present disclosure.

[0024]FIG. 4A is an isometric view illustrating a seed transport
subsystem (absent system support structure), of the seed sorter system
shown in FIG. 1, including a transport carousel having one of a plurality
of rotary vacuum cup banks mounted thereto, in accordance with various
embodiments of the present disclosure.

[0025]FIG. 4B is an isometric view of a rotary vacuum cup included in the
rotary vacuum cup bank shown in FIG. 4A, in accordance with various
embodiments of the present disclosure.

[0026]FIG. 5A is an isometric view of an imaging station (absent system
support structure), of the seed sorter system shown in FIG. 1, in
accordance with various embodiments of the present disclosure.

[0027]FIG. 5B is an exemplary schematic illustrating a 360° plane
in which the orientation of seed tips of a plurality of seeds are
determined at the imaging station shown in FIG. 5A.

[0028]FIG. 6A is an isometric view of a seed orientation station (absent
system support structure), of the seed sorter system shown in FIG. 1, in
accordance with various embodiments of the present disclosure.

[0029]FIG. 6B is an exemplary schematic illustrating a 360° plane
in which the seeds are controllably rotated at the seed orientation
station, shown in FIG. 6A, such that the seed tips of each seed have a
desired orientation.

[0030]FIG. 6c is an isometric partial view of the seed orientation
station shown in FIG. 6A, illustrating a seed purge hopper, in accordance
with various embodiments of the present disclosure.

[0031]FIG. 7 is an isometric view of the seed sampling system, shown in
FIG. 1, illustrating a seed sample and sort station, in accordance with
various embodiments of the present disclosure.

[0032]FIG. 8A is an isometric view of a seed sampling subsystem (absent
system support structure), of the seed sample and sort station, shown in
FIG. 7, in accordance with various embodiments of the present disclosure.

[0033]FIG. 8B is an isometric view of the seed grip and chip assembly of
the seed sampling subsystem, shown in FIG. 8A, in accordance with various
embodiments of the present disclosure.

[0034] FIG. 8C is an isometric view of an exemplary seed gripping finger
included in the seed grip and chip assembly, shown in FIG. 8B, in
accordance with various embodiments of the present disclosure.

[0035] FIG. 8D is a top view of a seed gripping mechanism of the seed grip
and chip assembly, shown in FIG. 8B, in accordance with various
embodiments of the present disclosure.

[0036]FIG. 8E is an isometric view of an exemplary cutting wheel of the
seed grip and chip assembly, shown in FIG. 8B, in accordance with various
embodiments of the present disclosure.

[0037]FIG. 9A is a front view of a seed and sample sorting subsystem, of
the seed sample and sort station, shown in FIG. 7, in accordance with
various embodiments of the present disclosure.

[0038]FIG. 9B is a side view of seed and sample sorting subsystem shown
in FIG. 9A.

[0039] FIG. 9C is diagonal view of a sample extraction nozzle manifold of
the seed and sample sorting subsystem shown in FIGS. 9A and 9B, in
accordance with various embodiments of the present disclosure.

[0040] FIG. 9D is an isometric view of a sample tray platform and X-Y
translation stage included in the seed and sample sorting subsystem shown
in FIGS. 9A and 9B.

[0041] FIG. 9E is an isometric view of a seed tray platform and X-Y
translation stage included in the seed and sample sorting subsystem shown
in FIGS. 9A and 9B.

DETAILED DESCRIPTION

[0042] The following description is merely exemplary in nature and is in
no way intended to limit the present teachings, application, or uses.
Throughout this specification, like reference numerals will be used to
refer to like elements.

[0043] FIGS. 1 and 2 illustrate an automated seed sampling system 10, in
accordance with various embodiments of the present disclosure. The seed
sorter system 10 includes a seed loading station 100, a seed transport
subsystem 200, a seed imagining station 300, a seed orientation station
400, a seed sampling and sort station 500 and a central controller system
(CCS) 700 that controls the operation of the seed sorter system 10.

[0044] Generally, the seed sampling system 10 is structured and operable
to repetitiously separate a select number of seeds, e.g., sets of eight
seeds at a time, from a bulk of seeds within a bulk seed hopper 104
(e.g., FIG. 3A, etc.) at the seed loading station 100. Additionally, the
seed sampling system 10 is structured and operable to image each set of
seeds at the imaging station 300. The images collected at the imaging
station 300 can be any desirable type of images. For example, the images
can be visual images, near infra-red (NIR) images or NMR/MRI images, or
any other type images. In various embodiments, the imaging station 300
collects at least one digital image of each set of seeds. The image data
collected of each set of seeds is communicated to a CCS 700 where the
image data is analyzed to determine the orientation, e.g., `tip out` or
`crown out`. The seed sampling system 10 is further structured and
operable to orient each set of seeds in a desired orientation, based on
the images of each respective set of seeds, at the orientation station
400. The seed sampling system 10 is still further structured and operable
to extract a sample (e.g., a tissue sample, etc.) from a selected area,
e.g., the crown, of each seed in each set of seeds. Further yet, the seed
sampling system 10 is structured and operable to then collect the
extracted samples in a plurality of sample trays 14 and sort the
respective sampled seeds into a plurality of seed trays 18 at the seed
sample and sort station 500.

[0045] Once a set of seeds is separated from the bulk of seeds at the seed
loading stations, as described below, the seeds are loaded onto one of a
plurality of rotary vacuum cup (RVC) banks 204. The respective set of
seeds is then sequentially positioned adjacent each of the imaging
station 300, the orientation station 400 and the sample and sort station
500, via the seed transport subsystem 200. More specifically, the seed
transport subsystem 200 includes an automated transport carousel 208 to
which the plurality of the RVC banks 204 are mounted. The automated
transport carousel 208 (FIG. 4A) is driven by a motor (not shown), e.g.,
a stepper motor, that incrementally rotates the transport carousel 208 to
sequentially advance each RVC bank 204 to each of the stations 100, 300,
400 and 500. Therefore, each set of seeds is retained by a respective RVC
bank 204 and transported to positions adjacent each of the imaging
station 300, the orientation station 400 and the sample and sort station
500 by the incremental rotation of the transport carousel 208.

[0046] The operation of the seed sorter system 10 is generally completely
controlled and automated by the CCS 700 such that the operations
performed by the imaging station 300, the orientation station 400 and the
sample and sort station 500 occur substantially without need for human
interaction, intervention or control. However, such actions as loading
the seeds into the bulk seed hopper 104 and/or physically manipulating
and/or changing the sample trays 14 and seed trays 18 (either
individually or collectively) can be performed manually with human
participation.

[0047] Generally, the CCS 700 includes one or more processors and/or
microprocessors, and one or more electronic data storage devices utilized
to store and execute various custom programs, applications and/or
algorithms to effectuate the operation of the seed sorter system 10.
Accordingly, the CCS 700 can comprise a specially programmed computer, or
computer system, in communication with associated system devices (not
shown) that enable communication with and control over the operations of
the various stations 100, 300, 400 and 500 and the transport subsystem
200 of the seed sorter system 10. Although the CCS 700 is exemplarily
illustrated in FIG. 2 as a single unit, the CCS 700 can be a single
computer based system or a plurality of computer based subsystems
networked together to coordinate the simultaneous operations of the seed
sorter system 10, as described herein. For example, in various
embodiments, the CCS 700 can include a main controller subsystem
networked together with a plurality of peripheral controller subsystems
(not shown), e.g., a peripheral controller subsystem for each station
100, 300, 400, 500 and transport subsystem 200. Each peripheral
controller subsystem can include one or more processors, microprocessors
and electronic data storage devices that effectuate communication with
various seed sorter system components, e.g., sensors, devices,
mechanisms, motors, tools, etc., and together with the main controller
subsystem cooperatively operate all the stations, systems and subsystems
of the seed sampler system 10. Or alternatively, the CCS 700 can comprise
a single computer communicatively connected to all the various system
components to cooperatively operate all the stations, systems and
subsystems of the seed sampler system 10.

[0048] As described above, the CCS 700 communicates with various seed
sorter system components that include various system sensors. The system
sensors operate to detect conditions of interest during operation of the
seed sorter system 10 and communicate that information to the CCS 700.
With this information, the CCS 700 generates control commands that
effectuate the operations and actions taken by the various stations and
components of the seed sorter system 10. For example, a sensed condition
can concern: the successful isolation of sets of seeds from the seed
hopper 104; the successful retention, or loading, of the each of the
seeds by a respective RVC bank 204; the proper positioning of each loaded
bank of seeds adjacent each respective station 300, 400 and 500; the
status (for example, position, location, vacuum, pressure, and the like)
of various component parts of the various stations 100, 300, 400 and 500;
operation, maintenance, performance, and error feedback from the various
components of each station 100, 300, 400 and 500 (separate from, or
perhaps comprising or in conjunction with, collected data); and the like.
More specifically, sensor information that is collected and processed for
use in controlling operation of the seed sorter system 10 can include
such information as: device or component status; error signals; movement;
stall; position; location; temperature; voltage; current; pressure; and
the like, which can be monitored with respect to the operation of each of
the stations, subsystems and associated components of the seed sorter
system 10.

[0049] It should be understood that the seed sorter system 10, as shown
and described herein, includes various stationary braces, beams,
platforms, pedestals, stands, etc., to which various components, devices,
mechanisms, systems, subsystems, assemblies and sub-assemblies described
herein are coupled, connected and/or mounted. Although such braces,
beams, platforms, pedestals, stands, etc., are necessary to the
construction of the seed sampler system 10, description of their
placement, orientation and interconnections are not necessary for one
skilled in the art to easily and fully comprehend the structure, function
and operation of the seed sampler system 10. Particularly, such braces,
beams, platforms, pedestals, stands, etc., are clearly illustrated
throughout the figures and, as such, their placement, orientation and
interconnections are easily understood by one skilled in the art.
Therefore, for simplicity, such braces, beams, platforms, pedestals,
stands, etc. will be referred to herein merely as system support
structures, absent further description of their placement, orientation
and interconnections.

[0050] Referring now to FIGS. 3A, 3B and 3C in various embodiments, the
seed loading station 100 includes the seed hopper 104 and a separating
wheel 108. The separating wheel 108 is mounted for rotation in a vertical
plane such that a portion of the separating wheel 108 extends into an
interior reservoir of the seed hopper 104. Another portion of the
separating wheel 108 extends outside of the seed hopper 104 such that a
face 110 of the separating wheel 108 is positioned adjacent a seed
collector 114. The seed separating wheel 108 includes a plurality of
spaced apart recessed ports 118 that extend through the face 110 and are
communicatively coupled to a vacuum system (not shown) such that a vacuum
can be provided at each of the recessed ports 118.

[0051] To initiate operation of the seed sampler system 10, seeds to be
sampled and tested are placed in the seed hopper 104 interior reservoir
and a vacuum is provided to at least some of the recessed ports 118,
e.g., the recessed ports 118 in the face 110 of the portion of the
separating wheel 108 extending into the interior reservoir of the seed
hopper 104. The seed separating wheel 108 is then incrementally rotated,
via an indexing motor 122, such that recessed ports 118 sequentially
rotate through the interior reservoir of the seed hopper 104, out of the
seed hopper 104, and past seed collector 114 before re-entering the
interior reservoir of the seed hopper 104. As the separating wheel
incrementally rotates and the recessed ports 118 incrementally pass
through the seed hopper 104 interior reservoir, individual seeds are
picked up and held at each recessed port 118 by the vacuum provided at
the respective recessed ports 118. As the separating wheel 108
incrementally rotates, the seeds are carried out of the seed hopper 104
to the seed collector 114 where each seed is removed from the face 110 of
the separating wheel 108.

[0052] In various embodiments, the seed collector 114 includes a wiper
(not shown) that physically dislodges each seed from the respective
recessed port 118 as the separating wheel 108 incrementally rotates past
the seed collector 114. Alternatively, in various other embodiments, each
seed can be released from respective recessed port 118 by temporarily
terminating the vacuum at each individual recessed port 118 as the
individual recessed port 118 is positioned adjacent the seed collector
114. In still other embodiments, each seed can be blown from the
respective recessed port 118 by temporarily providing forced air at each
individual recessed port 118 as the individual recessed port 118 is
positioned adjacent the seed collector 114.

[0053] After each seed is removed from the separating wheel 108, the seed
is funneled into one of a plurality of first transfer tubes 126 having
their proximal ends connected to openings 128 in a tube shuttle 130. The
tube shuttle 130 is mounted to a carriage 134 that is movably mounted to
a linear translation stage 138 that includes an actuator 142 controllable
by the CCS 700 to bi-directionally move the carriage 134, tube shuttle
130 and proximal ends of the first transfer tubes 126 along the
translation stage 138. Therefore, as each seed is removed from the
separating wheel 108, the seed is funneled into one of the first transfer
tubes 126. Then the CCS 700 moves the tube shuttle 130 along the
translation stage such that a subsequent first transfer tube 126 will
receive the next seed removed from the separating wheel 108. This process
of removing seeds is repeated until a seed has been deposited into each
of the first transfer tubes 126. As each seed is removed from the
separating wheel 108 and deposited into a first transfer tube 126, each
seed passes through the respective first transfer tuber 126, via gravity,
vacuum or forced air, to a queuing stack 150.

[0054] As shown in FIG. 3B, the queuing stack 150 includes a plurality of
upper chambers 154, e.g., eight upper chambers 154. A distal end of each
first transfer tube 126 terminates at a corresponding one of the upper
chambers 154. Each upper chamber 154 includes an automated upper release
mechanism 156, e.g., a flapper gate, that, under control of the CCS 700,
retains the respective seed within the upper chamber 154. Once each upper
chamber has a seed deposited therein, the upper release mechanisms 156
are commanded to release the seeds into a plurality, e.g., eight, of
corresponding lower chambers 158. Similar to the upper chambers 154, each
lower chamber 158 includes an automated lower release mechanism 160,
e.g., a flapper gate, that, under control of the CCS 700, retains the
respective seed within the lower chamber 154. The lower chambers 158
retain the seeds until such time as the CCS 700 commands the lower
release mechanisms 160 to release the seeds into a plurality of
corresponding second transfer tubes 162 having their proximal ends
connected to the lower chambers 158.

[0055] As shown in FIG. 3C, a distal end of each second transfer tube 162
terminates at a corresponding one a plurality of elevator chambers 164,
e.g., eight, of an elevator hopper 166. Thus, as the seeds are released
from the lower chambers 158 into the second transfer tubes 162, each seed
passes through the respective second transfer tube 162, via gravity,
vacuum or forced air, into a corresponding one of the elevator chambers
164. Once a seed is deposed into each elevator chamber 164, the group of
seeds therein constitutes a set of seeds, as used herein. The elevator
hopper 166 additionally includes a plurality of elevator piston rods 170.
Each elevator piston rod 170 is positioned within, and extendable
through, an aperture 174 formed in a funnel-shaped bottom 178 of a
corresponding elevator chamber 164. In various embodiments, a distal end
of each elevator piston rod 170 is formed to have a concave recess 180
shaped to cradle the seeds received from the lower chambers 158 of the
queuing stack 150. Additionally, the angled sides of the funnel-shaped
bottom 178 allows for the seeds entering the respective elevator chambers
164 to fall onto their sides, i.e., lay flat, and be centered within the
recess 180 of the respective elevator piston rods 170.

[0056] Each elevator piston rod 170 can be extended and retracted through
the respective elevator chamber aperture 174 by a corresponding one of a
plurality of piston actuators 182, as controlled by the CCS 700. More
particularly, prior to the seeds being deposited in the elevator chambers
164, the CCS 700 commands the piston actuators 182 to retract the piston
rods 170 to a retracted position where the recessed distal ends of each
piston rod 170 is substantially flush with the bottom of each respective
elevator chamber 164 within the respective aperture 174, as exemplarily
illustrated by the six leftmost piston rods 170 in FIG. 3C. Then once the
seeds are deposited into the elevator chambers 164, the seeds are
retained and cradled within the respective piston rod recesses 180, until
such time as the CCS 700 commands the piston actuators to extend the
piston rods 170 to an extended position, as exemplarily illustrated in
phantom by the two rightmost piston rods 170 in FIG. 3C. Extending the
piston rods 170 raises each respective seed out of the respective
elevator chamber 164 to a cued position where the set of seeds are
presented for removal, processing and sampling by a RVC bank 204, as
describe below.

[0057] Referring now to FIGS. 4A and 4B, as described above, the seed
transport subsystem 200 includes a plurality of RVC banks 204 mounted to
the transport carousel 208. For simplicity and clarity, FIG. 4A
illustrates a single RVC bank 204 mounted to the transport carousel 208,
however, it should be understood that seed transport subsystem 200
includes a plurality of RVC banks 204 mounted thereto. For example, in
various embodiments, the seed transport subsystem 200 includes four RVC
banks 204, whereby each RVC bank 204 is mounted to one of four sides of
the transport carousel 208. Each RVC bank includes a plurality of rotary
vacuum cup (RVC) devices 212 controllable by the CCS 700 to remove a set
of cued seeds from the elevator piston rods 170 and sequentially
transport the respective seed set to each of the imaging station 300, the
orientation station 400 and the sample and sort station 500.

[0058] Each RVC device 212 includes a vacuum cup 216 mounted to a first
end of a rotary shaft 220, and a friction plate 224 mounted to an
opposing second end of the rotary shaft 220. Each RVC device 212
additionally includes a shaft actuator 228 controllable by the CCS 700 to
bidirectionally move the shaft 220, vacuum cup 216 and friction plate 224
along the longitudinal axis of the rotary shaft 220. That is, each
actuator 228 is controlled by the CCS 700 to raise and lower the
respective vacuum cup 216 as needed throughout operation of the seed
sorter system 10. Each vacuum cup 216 is communicatively connected to a
vacuum source (not shown) that is controlled by the CCS 700 to
selectively provide a vacuum at a tip 232 of each vacuum cup 216. Each
RVC device 212 further includes a biasing device 236, e.g., a spring,
configured to apply a constant force on the rotary shaft 220 in the X
direction. The force applied in the X direction by the biasing devices
236 maintains a locking mechanism (not shown) of each respective RVC
device 212 engaged. Engagement of the locking device prevents angular
rotation of the respective rotary shaft 220 and vacuum cup 216 about the
longitudinal axis of the rotary shaft 220 until the locking mechanism is
disengaged, as described below.

[0059] In coordination with a set of seeds being loaded into the elevator
chambers 164, the CCS 700 positions an empty RVC bank 204, i.e., an RVC
bank 204 without a set of seeds retained by the respective vacuum cups
216, above the elevator bank 166. The RVC devices 212 are located and
mounted to the transport carousel 208, and the motor of the transport
carousel 208 is controlled, such that when an RVC bank 204 is positioned
adjacent the loading station 100, the vacuum cup 216 of each RVC device
212 is positioned directly above a corresponding one of the elevator
chambers 164. More particularly, when an RVC bank 204 is positioned
adjacent the loading station 100, the vacuum cup 216 of each RVC device
212 is positioned directly above the elevator piston rod 170 of the
corresponding elevator chamber 164. Once the elevator chambers 164 are
loaded with a set of seeds, and an empty RVC bank 204 is positioned
adjacent the loading station 100, the CCS 700 can command the elevator
piston rods 170 to raise the set of seeds to the cued position. The RVC
devices 212 are further located and mounted to the transport carousel
208, such that when the elevator piston rods 170 are in the extended
position, i.e., the set of seeds are cued, each seed is in light contact
with, or close proximity to, the corresponding vacuum cup 216. A vacuum
is then provided to each vacuum cup tip 232. The vacuum cup tips 232 are
sized, and fabricated from a suitable material, such that when the vacuum
is provided, each respective seed is firmly retained on the respective
tip 232. The CCS 700 then retracts the elevator piston rods 170 leaving
the set of seeds firmly retained on the respective vacuum cup tips 232.
The retained set of seeds can then be positioned adjacent the imaging
station 300, via advancement of the transport carousel 204.

[0060] Referring now to FIGS. 5A and 5B, in various embodiments, the
imaging station 300 includes at least one imaging device 304 mounted to
system support structure such that the one or more imaging devices 304
is/are positioned under the RVC bank 204 and the respective set of seeds
when the set of seeds is advanced from the loading station 100. In
various embodiments, the imaging station 300 includes a first imaging
device 304 positioned and operable to collect image data for a first
one-half of the seed set, and a second imaging device 304 positioned and
operable to collect image data for a second one-half of the seed set.
More particularly, the first imaging device 304 is mounted to the system
support structure such that a field of view of the first imaging device
304 includes a bottom side of a first half of the seeds positioned
adjacent the imaging station 300. And, the second imaging device 304 is
mounted to the system support structure such that a field of view of the
second imaging device 304 includes a bottom side of a second half of the
seeds positioned adjacent the imaging station 300.

[0061] As used herein, reference to the bottom side of the seeds refers to
the side of the seeds that is facing downward with respect to the
orientation of each seed as retained by the respective vacuum cup 216. As
described above, the shape of the elevator chamber bottoms 178 and the
shape of the recesses 180 at the distal ends of each elevator rod 170 are
designed such that each seed is preferably retained on the vacuum cup
tips 232 by one of the opposing broad sides of each respective seed. That
is, each seed is preferably held on the respective vacuum cup 216 by one
of the broader sides such that germ of the seed is viewable by the
imaging device(s) 304 and the tip of each seed is pointing anywhere
within a 360° plane that is substantially orthogonal to the
respective vacuum cup 216. The imaging device(s) 304 may be any suitable
imaging device selected in accordance with the imaging goals of seed
sorter system 10. For example, in connection with an analysis for
external seed coat, the first imaging device 304 may comprise a digital
camera operable in the visible light range. Alternatively, for internal
seed analysis, the first imaging device 304 may comprise a camera
operable in the near infra-red light range (see, U.S. application for
patent Ser. No. 09/698,214, the disclosure of which is hereby
incorporated by reference). Still further, the first imaging device 304
may comprise a camera which implements NMR/MRI imaging techniques (see,
U.S. application for patent Ser. No. 09/739,871, the disclosure of which
is hereby incorporated by reference).

[0062] The imaging station 300 further includes a light source 312 mounted
to system support structure for illuminating the field of view of the
imaging device(s) 304. The source 312 can be any type of light source
suited for the particular imaging application of the seed sorter system
10. For example, the light source 312 can be one or more incandescent
lights, fluorescent lights, ultraviolet lights, infrared lights, etc. In
various embodiments, the light source 312 comprises a bank of light
emitting diodes (LEDs), e.g., 630 nm LEDs. For example, the light source
312 comprises a bank of LEDs wherein each seed in the seed set has a
corresponding LED as the primary light source illuminating the respective
seed. Additionally, in various embodiments, each vacuum cup 216 includes
a dark colored, e.g., black, background disk 240 (FIG. 4B) that provides
a dark background for each seed during imaging and prevents image data
interference from system components and structure beyond the seeds and
within the field of view of imaging device(s) 304.

[0063] The image data is transmitted to the CCS 700 and stored (at least
temporarily) in an electronic data storage device of the CCS 700. The CCS
700 analyzes the data to determine a directional orientation of the tip
of each seed. That is, the CCS 700 analyzes the image data to determine
which direction the tip of each individual seed is pointing within the
360° plane substantially orthogonal to the respective vacuum cup
216. For example, with reference to FIG. 5B, if a point on the
360° plane that is directly opposite the transfer carousel 208 is
considered the origin, i.e., 0°, the CCS 700 may analyze the image
data to determine that the tip one of the seeds in the seed set is
oriented at 90°, while the tip of another of the seeds in the seed
set is oriented at 315°, and the tip of yet another seed is
oriented at 200°, etc. Once the image data of the respective seed
set is collected and transmitted to the CCS 700, by the imaging device(s)
304, the transfer carousel 208 is advanced to position the seed set
adjacent the orientation station 400.

[0064] Referring now to FIGS. 6A and 6B, in various embodiments, the
orientation station 400 includes motor bank 404 that includes a plurality
of rotary motors 408, e.g., a number of rotary motors 408 equal to the
number of RVC 212, independently controlled by the CCS 700. In some
embodiments, the rotary motors 408 comprise stepper motors. Each motor
408 includes a rotary shaft 412 having a clutch plate 416 mounted to a
distal end thereof. The motor bank 404 is mounted to a linear actuator
420, e.g., a pneumatic slide, that is mounted to system support structure
such that when the RVC bank 204 is positioned adjacent the orientation
station 400, each motor 408 is positioned directly above a respective one
of the RVC devices 212. More specifically, when the RVC bank 204 is
positioned adjacent the orientation station 400, the clutch plate 416 of
each motor 408 is positioned directly above, and in alignment with, a
respective one of the RVC friction plates 224.

[0065] Once the imaged set of seeds is advanced to the orientation station
400, the actuator 420 lowers the motor bank 404 such that the clutch
plates 416 of each motor 408 engage the corresponding friction plates 224
of the respective RVC device 212. Additionally, the actuator 420 is
lowered such that the clutch plates 416 apply force to each friction
plate 224 in the Y direction that overcomes the force in the X direction
applied by the RVC biasing devices 236. Accordingly, each RVC friction
plate 224, rotary shaft 220 and vacuum cup 216 is moved downward, thereby
disengaging the RVC locking mechanism and allowing each RVC friction
plate 224, rotary shaft 220 and vacuum cup 216 to rotate. Then, based on
the analyzed image data collected at the imaging station 300, each motor
408 is independently controlled by the CCS 700 to rotate the respective
friction plates 224 and corresponding vacuum cups 216 to independently
properly orient each respective seed for sampling at the sample and sort
station 500, as described below. More particularly, based on the analyzed
image data for each independent seed, each motor 408 is independently
controlled to rotate the respective seed such that the tip of the seed is
oriented approximately at 0°. More importantly, each seed is
independently rotated, if necessary, to position the cap of the seed at
approximately 180° such that a sample can be removed from that cap
of each seed at the sample and sort station 500, as described below.

[0066] Once each seed of the set is oriented with the cap of each
respective seed oriented, or positioned, at approximately 180°,
the CCS 700 commands the actuator 420 to raise the motor bank 404 to
disengage the clutch plates 416 from the friction plates 224. As the
motor bank 404 is raised and the clutch plates 416 are disengaged from
the friction plates 224, the biasing devices 236 of each RVC device 212
move each respective rotary shaft in the X direction thereby engaging
each respective locking device. Thus, each rotary cup 216 and
corresponding seed held thereon, is maintained in the orientation with
the seed cap at approximately the 180° position, as illustrated in
FIG. 6B. The CCS 700 then advances the transfer carousel 208 to position
the properly oriented seed set adjacent the sample and sort station 500.

[0067] Referring now to FIG. 6c, in various embodiments, the orientation
station 400 further includes a seed purge hopper 424 for receiving the
set of seeds held by the respective RVC bank 204. The seed purge hopper
424 is mounted to system support structure such that a trough 428 of the
seed purge hopper 424 is positioned under the vacuum cups 216 of the
respective RVC bank 204 for receiving seeds discharged from the
respective vacuum cups 216. More specifically, the seed purge hopper 424
can be utilized to offload all the seeds held by each RVC bank 204 of the
seed transport subsystem 200. To offload all the seeds, each RVC bank 204
is sequentially advanced to the orientation station 400 at which time the
vacuum source being supplied to each respective vacuum cup 216 is
terminated. When the supplied vacuum is terminated, the seeds are
released from the vacuum cups 216 and fall into the seed purge hopper
trough 428 where they can be collected and returned to the bulk seed
hopper 104 at a later time. Thus, if operation of the seed sorting system
10 needs to be terminated, all the seeds held by the RVC banks 204 can be
purged and collected.

[0068] In various embodiments, the seed sorting system 10 includes an
emergency stop button 22, shown in FIG. 1, for stopping and shutting down
the seed sorting system 10. For example, in the case of an emergency, the
emergency stop button 22 can be depressed and the all operation of the
seed sorting system 10 would cease. Also, in various embodiments, the
seed sorting system 10 includes a system pause button 26, shown in FIG.
1, for temporarily pausing operation of the seed sorting system 10. For
example, if a jam occurred in one of the first transfer tubes 126 of the
seed loading station 100 such that one or more RVC banks did not `pick
up` a full set of seeds, the system pause button could be depressed to
pause operation of the seed sorting system 10. In the paused state, the
vacuum source can remain actuated such that all seeds are retained by the
respective RVC vacuum cups 216 until such time as the seeds are purged
into the seed purge hopper 424 or operation is reinitiated and the seeds
are sampled, as described below.

[0069] Referring now to FIG. 7, in various embodiments, the seed sample
and sort station 500 includes a seed sampling subsystem 510 and a seed
and sample sorting subsystem 570. The seed sampling subsystem 510 is
controllable by the CCS 700 to extract a sample from each seed in the
respective seed set positioned adjacent the seed sample and sort station
500. The seed and sample sorting subsystem 570 is additionally
controllable by the CCS 700 to sort the sampled seeds to the seed trays
18 and sort the corresponding seed sample to the sample trays 14 while
tracking and mapping the locations of the corresponding sampled seeds and
seed samples in the respective seed and sample trays 18 and 14. The
locations of the seed samples and the locations of the corresponding
sampled seeds in the trays 14 and 18 are matched so that the sampled seed
may later be correlated to the sample taken therefrom (e.g., after
analysis of the sample, etc.).

[0070] Referring now to FIGS. 8A, 8B, 8C and 8D, in various embodiments,
the seed sampling subsystem 510 includes a plurality of seed grip and
chip assemblies 512, e.g., a number of seed grip and chip assemblies 512
equal to the number of RVC devices 212 included in each RVC bank 204. The
seed sampling subsystem 510 additionally includes a press plate bank 514
mounted to a linear actuator 516, e.g., a pneumatic slide. The press
plate bank 514 includes a plurality of press plates 518 fixedly mounted
to a press plate bank header 520 that is coupled to the linear actuator
516. The actuator 516 is mounted to system support structure such that
when the RVC bank 204 is positioned adjacent the sample and sort station
500, each press plate 518 is positioned directly above a respective one
of the RVC devices 212. More specifically, when the RVC bank 204 is
positioned adjacent the orientation station 400, each press plate 518 is
positioned directly above, and in alignment with, a respective one of the
RVC friction plates 224.

[0071] Once the oriented set of seeds is advanced to the sample and sort
station 500, the actuator 516 lowers the push plate bank 514 such that
the push plates 518 engage the corresponding friction plates 224 of the
respective RVC device 212. As the actuator 420 is lowered, the push
plates 518 apply force to each friction plate 224 in the Y direction that
overcomes the force in the X direction applied by the RVC biasing devices
236. Accordingly, each RVC friction plate 224, rotary shaft 220 and
vacuum cup 216 is moved downward, thereby disengaging the RVC locking
mechanism. However, since the press plates 518 are fixedly mounted to the
header 520, each RVC friction plate 224, rotary shaft 220 and vacuum cup
216 can not rotate and each seed remains properly oriented as it is moved
downward in the Y direction.

[0072] With particular reference to FIGS. 8B and 8D, in accordance with
various embodiments, each grip and chip assembly 512 includes a seed
gripping mechanism 522 and a sample extraction mechanism 524. Although,
each grip and chip assembly 512 is independently controlled by the CCS
700, the structure and function for each grip and chip assembly 512 is
substantially identical. Therefore, the structure and function of the
plurality of grip and chip assemblies will be described herein with
reference to a single grip and chip assembly 512. The seed gripping
mechanism 522 is operable, as controlled by the CCS 700, to firmly hold
each respective seed as the sample extraction mechanism 524 removes a
portion, i.e., a sample, of the seed coat and inner seed material from
the crown of the respective seed. The extracted sample can then be
utilized to test and analyze the various traits of the respective seed.
Importantly, the sample is extracted from the crown in a non-destructive
manner such that germination viability of the seeds can be preserved.

[0073] In various embodiments, the seed gripping mechanism 522 includes an
actuator 526, e.g., a pneumatic clamp, that is controllable by the CCS
660 to bidirectionally move a pair of opposing actuator arms 528 toward
and away from each other, i.e., open and close the actuator arms 528. For
example, in various embodiments, the actuator 526 is operable to move the
opposing actuator arms 528 toward and away from each other along the line
M (FIG. 8B). The actuator arms 528 are structured to removably retain a
pair of opposing seed gripping fingers 530 structured to firmly hold the
respective seed as the sample is extracted by the sample extraction
mechanism 524, as described below. An exemplary gripping finger 530 is
illustrated in FIG. 8C. Each gripping finger 530 includes a head 532
having a contoured face 534. The face 534 can be shaped or formed to have
any conformation suitable for firmly and steadily holding the respective
seed as the sample is extracted. In various embodiments, the face 534 is
particularly designed to have a wedge-like conformation such that as the
actuator 526 closes gripping fingers 530 around the seed, the seed is
forced toward a cutting wheel 540 of the sample extraction mechanism 524
and into abutment with a justification block 562 of the sample extraction
mechanism 524. The justification block 562 includes a cutting wheel guide
slot 533 that allows the cutting wheel 540 access to seed. Thus, the
gripping fingers 530 firmly and justification block 562 hold the seed on
three sides and prevent the respective seed from moving in a direction
away from the sample extraction mechanism 524 as the sample is being
extracted. In various embodiments, the gripping finger head 532 is
connected to, or integrally formed with, a mounting post 536 structured
to fit within, or mate with, a mounting hole 538 in each actuator arm
528.

[0074] When the RVC bank 204 and properly oriented seed set are advanced
to the seed sample and sort station 500, the CCS 700 commands the seed
gripping actuator 526 to open the actuator arms 528 such that the
gripping finger faces 534 have a space between them large enough to allow
a seed to be easily positioned therebetween. The CCS 700 then commands
the press plate bank actuator 516 to lower the press plate bank 514 to
engage the press plates 518 with the friction plates 224. More
particularly, the force on the friction plates 224 moves the vacuum cups
216 and respective seed downward toward a sampling position, i.e., the
gap between gripping fingers 530. Each grip and chip system 512 is
mounted to system support structure such that each sampling position, or
gap, between the gripping finger faces 534 is precisely aligned below the
respective vacuum cup tip 232. Thus, when the press plate bank actuator
516 pushes the friction plates 224, vacuum cups 216 and oriented seeds
downward, the oriented seeds are moved to the sampling positions between
the gripping fingers 530 of the respective seed gripping mechanism 522.

[0075] The CCS 700 then commands the seed gripping actuator 526 to close
the actuator arms 528 such that the gripping finger faces 534 engage the
respective seed to firmly retain the seed without damaging the seed. Once
the seed is firmly retained between the gripping fingers 530, the push
plate bank actuator can be commanded to raise, and the vacuum provided at
the vacuum cup tip 232 terminated, to thereby release the respective
seed. Or, alternatively, the CCS 700 can maintain the vacuum cup 216 in
contact with the seed to provide additionally support for the seed as the
sample is being extracted.

[0076] The sample extraction mechanism 524 includes cutting wheel 540
rotationally mounted within a cutting wheel fixture 542 and rotationally
driven by a cutting wheel motor 544. Although the cutting wheel 540 is
shown in FIG. 8B to be belt driven by the cutting wheel motor 544,
alternatively the cutting wheel 540 can be shaft driven, chain driven,
direct gear driven, etc., by the cutting wheel motor 544 and remain
within the scope of the present disclosure. The cutting wheel 540 is
mounted on a shaft 546 that is rotationally mounted within the cutting
wheel fixture 542. Additionally, a drive wheel 548 is mounted to, or
formed with, the shaft and operatively coupled to the cutting wheel motor
544, for example, by a drive belt 550, such that actuation of the motor
544 will rotate the drive wheel 548, shaft 546 and cutting wheel 540
within the cutting wheel fixture 542. More specifically, the cutting
wheel 540 is mounted to the shaft 546 in a cam fashion, e.g., the shaft
546 can be an offset shaft, such that as the drive wheel 548 and shaft
546 are rotated by the motor 544, a peripheral cutting edge 552 of the
cutting wheel 540 rotates and progressively moves toward the seed
gripping mechanism 552, and specifically toward the seed retained between
the gripping fingers 530. The cutting edge 552 comprises an abrasive or
sharp-edged surface, e.g., a saw-toothed surface, that will remove the
seed coat and inner seed material from the crown of the respective seed.
Thus, as the cutting wheel 540 is rotated, the cutting edge 552 will
contact and begin to cut or abrade the seed crown. As the cutting wheel
540 continues to rotate, the cutting edge 552 will penetrate a desired
depth or distance into the crown, depending on the amount of angular
rotation of the cutting wheel 540, as controlled by the CCS 700. That is,
the greater the amount of angular rotation of the cutting the wheel 540,
the deeper the cutting edge 552 will penetrate into the seed crown and
the more sample that will be extracted.

[0077] The seed gripping mechanism 522 additionally includes a seed dump
bowl 554 (FIG. 8D) having a drain port 556 formed in the bottom of the
dump bowl 554. The dump bowl 554 is shaped to funnel a sampled seed into
the drain port 556 after a sample has been removed from the respective
seed and the seed is released from being held between the gripper fingers
530. A drain tube 558 (shown in FIGS. 9A and 9B) is connected to the
drain port 556 to transfer the released seed to one of the seed trays 18
positioned below the grip and chip assembly 512, as describe below. The
seed gripping mechanism 522 further includes a sample extraction orifice
560 located in the justification block 562, below the cutting wheel guide
533, such that cutting edge 552 and periphery portion of the cutting
wheel 540 extends over the sample extraction orifice 560. A sample
extraction tube 564 (FIGS. 9A and 9B) is connected to the sample
extraction orifice 560 and a vacuum is controllably provided to the
sample extraction tube 564 and thus, at the sample extraction orifice
560. As the cutting wheel 540 removes the sample from the respective
seed, the vacuum provided at the sample extraction orifice 560, via the
sample extraction tube 564, draws the sample into the sample extraction
orifice 560. The sample is then passed through the sample extraction tube
564 and deposited into one of the sample trays 14.

[0078] Therefore, once the seed is retained between the gripping fingers
530, the CCS 700 commands the cutting wheel motor 544 to angularly rotate
the drive wheel 548 through a predetermined angle and at a predetermined
rate of rotation. For example, the CCS 700 can command the cutting wheel
motor 554 to rotate the drive wheel 548 ninety-five degrees at thirty
revolutions-per-minute (RPMs). Accordingly, the cutting wheel 540 is
rotated through the predetermined angle, at the predetermined RPMs. As
the cutting wheel 540 rotates, the cam action of cutting wheel 540
mounting rotates and advances the cutting edge 552 toward and into the
seed, thereby removing a sample from the respective seed. As the sample
is removed, the vacuum at the sample extraction orifice 560 draws the
sample into the sample extraction tube 564 where the sample is
transferred to one of the sample trays 14. The CCS 700 then commands the
cutting wheel motor 544 to reverse the direction of rotation to withdraw
the cutting wheel 540 from the seed and return the cutting wheel to a
home position, ready to remove a sample from a subsequent seed.
Subsequent to, or substantially simultaneously with the withdrawal of the
cutting wheel 540, the CCS 700 commands the seed gripping mechanism 522
to release the sampled seed, allowing the seed to fall, via gravity,
vacuum or forced air, into the drain tube 556 to transfer the sampled
seed to one of seed trays 18.

[0079] Referring now to FIG. 8E, in various embodiments, the cutting wheel
540 is structured to have a saw-toothed cutting edge 552 that includes a
plurality of teeth 566. Moreover, each tooth 566 includes a lateral
cutting tip 568 that is formed to avoid movement, e.g., `chattering`, of
the seed being sampled and allow the seed to remain stationary within the
gripping fingers 530. For example, the lateral cutting tip 568 of each
tooth can have a specific angle α, e.g, a 60° angle, such
that as the cutting wheel 540 cuts through the respective seed, a leading
end of the cutting tip 568 of each subsequent tooth 566 engages the seed
before a trailing end of the cutting tip 568 of each preceding tooth 566
disengages the seed.

[0080] In various embodiments, each cutting wheel 540, i.e., each cutting
wheel motor 544, is independently controlled by the CCS 700, but each
cutting wheel 540 is commanded to have approximately the same rotational
speed and/or angular rotation. Therefore, when a set of seeds is held
within the seed gripping mechanisms 522, the respective cutting wheels
540 are each commanded to rotate through approximately the same angle of
rotation and at the same speed. Accordingly, the cam rotation of the
cutting wheels 540, as described above, will advance the respective
cutting wheels 540 approximately the same distance toward each of the
seed. Thus, smaller seeds may not have the same amount of sample
extracted as larger seeds. In such embodiments, the rotational speed
and/or amount of angular rotation for each cutting wheel 540 is
determined by empirical data and programmed into the CCS 700.

[0081] In various other embodiments, each cutting wheel 540, i.e., each
cutting wheel motor 544, is independently controlled by the CCS 700.
Therefore, the rotational speed and/or amount of angular rotation for
each independent cutting wheel 540 can be controlled and adjusted for
each seed positioned and held by the seed gripping mechanism 522 of each
respective grip and chip assembly 512. For example, the seed held within
a seed gripping mechanism 522 of a first grip and chip assembly 512 may
be smaller in size than a seed held within a seed gripping mechanism 522
of an adjacent second grip and chip assembly 512. In such a case, the
cutting wheel 540 of first grip and chip assembly 512 can be commanded to
have a greater angular rotation than the cutting wheel 540 of second grip
and chip assembly 512. Therefore, the cam rotation of the cutting wheels
540, as described above, will advance the cutting wheel 540 of the first
chip and grip assembly 512 further toward the smaller seed such that
approximately equal amounts of sample will be extracted from the smaller
seed as from the larger seed. Furthermore, in such embodiments, the
rotational speed and/or amount of angular rotation for each independent
cutting wheel is based on the imaging data collected for each respective
seed at the imaging station 300.

[0082] Referring now to FIGS. 9A, 9B, 9C, 9D and 9E, as described above,
as the sample is extracted from the respective seed, the sample is drawn
into the sample extraction tube 564. More specifically, the sample
extraction orifice 560 of each grip and chip assembly 512 has a first end
of a respective sample extraction tube 564 connected thereto, and a
second end of each respective sample extraction tube 564 is connected to
a sample extraction nozzle (SEN) manifold 572. The SEN manifold 572
includes a manifold block 574 to which the sample extraction tubes 564
are connected, and from which a plurality of exhaust tubes 578 extend,
e.g., a number of exhaust tubes 578 equal to the number of sample
extraction tubes 564 can extend from the manifold block 574. The SEN
manifold 572 additionally includes a plurality of discharge nozzles 580
that are in fluid communication with the extraction tubes 564. More
specifically, the manifold block 574 includes a number of bores or
passages (not shown), equal to the number of sample extraction tubes 564,
which extend through the manifold block 574. Each extraction tube 564 is
connected to a first end of a corresponding manifold block bore and a
corresponding one of the discharge nozzles 580 extends from an opposing
second end of each manifold block bore.

[0083] As most clearly illustrated in FIG. 9D, the seed and sample sorting
subsystem 570 further includes a sample tray platform 582 adapted to
securely retain a plurality of the sample trays 14 in fixed positions and
orientations. Each sample tray 14 includes a plurality of sample wells
30, each of which are adapted for receiving a sample extracted by one of
the grip and chip assemblies 512. For example, in various embodiments,
each sample tray 14 can be a ninety-six well tray. Moreover, the
discharge nozzles 580 extending from the SEN manifold 572 are spaced
apart and arranged to be congruent with the spacing and arrangement of
the wells 30 within the sample trays 14. The sample tray platform 582 is
mounted to an X-Y stage 584 that is a two-dimensional translation
mechanism, including a X axis translating track 586 and a Y axis
translating track 588. The X-Y stage 584 additionally includes a first
linear actuator 590 operable to bidirectionally move a first carriage
(not shown) along the length of the X axis translating track 586. The X-Y
stage 584 further includes a second linear actuator 592 operable to
bidirectionally move a second carriage (not shown) along the length of
the Y axis translating track 588. The Y axis translating track 588 is
mounted to the first carriage and the sample tray platform 582 is mounted
to the second carriage.

[0084] The SEN manifold 572 is connected to system support structure to
position the SEN manifold 572 above the X-Y stage 584 and the sample
platform 582 holding the plurality of sample trays 14. More particularly,
the SEN manifold 572 is mounted to a linear actuator 594, e.g., a
pneumatic slide, controllable by the CCS 700 to bidirectionally move the
SEN manifold 572 along the Z axis, e.g., up and down. The first and
second linear actuators 590 and 592 are controlled by the CCS 700 to
precisely move the sample tray platform 582 in two dimensions. More
particularly, the first and second actuators 590 and 592 move the sample
tray platform 582 within an X-Y coordinate system to precisely position
any selected group of adjacent wells 30 of any one or more selected
sample trays 14 at a target location directly beneath the SEN manifold
572.

[0085] In operation, prior to the grip and chip assemblies 512 extracting
samples from the respective seeds advanced to the seed sample and sort
station 500, the CCS 700 controls the X-Y stage 584 to position a
selected group of wells 30 at the target location. The CCS 700 then
commands the SEN manifold actuator 594 to lower the SEN manifold 572 to
position each of the discharge nozzles 580 in alignment with and in close
proximity to, or in contact with, a corresponding one of the wells 30.
Once the selected group of wells 30 is positioned at the target location
and the SEN manifold 572 is lowered, the CCS 700 commands the grip and
chip assemblies 512 to extract the samples from the respective seeds.
Each sample is drawn into a corresponding sample extraction tube 564, as
described above, and the vacuum provided to each sample extraction tube
564 transfers each sample through the respective sample extraction tube
564 to the corresponding discharge nozzle 580. Each seed is then
discharged into the corresponding sample tray wells 30. The SEN manifold
actuator 594 then raises the SEN manifold 572, a subsequent group of
wells 30 is positioned at the target position, and the SEN manifold 572
is lowered in preparation for a subsequent set of samples to be extracted
and deposited into the wells 30.

[0086] In various embodiments, each discharge nozzle 580 includes a seal
596 that contacts the sample tray(s) 14 and creates a seal between each
discharge nozzle 580 and the corresponding well 30 when the SEN manifold
572 is lowered. Thus, the seals 596 ensure that substantially all the
sample being discharged from each discharge nozzle 580 is deposited into
the corresponding well 30 without cross-contamination by adjacent samples
escaping around the discharge nozzles 580. The seals 596 can be any seal
suitable for creating a seal between each discharge nozzle 580 and the
corresponding well 30, e.g., an O-ring, gasket or bushing.

[0087] As the sample trays 14 are placed on the sample tray platform 582,
a tray identification number, e.g., a bar code, for each sample tray 14
and the location of each sample tray 14 on the platform 582 is recorded.
Additionally, as each extracted sample is deposited into a well 30, an
X-Y location of the well 30 on the sample tray platform 582 is recorded.
The recorded tray and well positions on the sample tray platform 582 can
then be compared to the X-Y locations of each deposited extracted sample,
to map the specific extracted sample in each well 30 of each sample tray
14. In various embodiments, the sample tray platform 582 is removably
coupled to the X-Y stage 584 such that one or more sample tray platforms
582 can be loaded with the sample trays 14 offline, i.e., away from the
seed sorter system 10, and conveniently coupled to and decoupled from the
X-Y stage 584.

[0088] Additionally, in various embodiments, the extraction tubes 564 are
fabricated from static dissipative tubing so that a portion of the
extracted samples do not stick to the inside walls of the extraction
tubes 564 and cause cross-contamination of the samples. Furthermore, in
various embodiments, the seed and sample sorting subsystem is structured
and operable to `blow out` the extraction tubes 564 and discharge nozzles
580 between cycles. Therefore, any sample residue accumulated in the
extraction tubes 564 and discharge nozzles 580 is cleaned out between
cycles. For example, air pressure can be drawn or forced through the
extraction tubes 564 and discharge nozzles 580 and exhausted into the
exhaust tubes 578. The exhaust tubes 578 can be coupled to an exhaust
manifold 598 that carries any residual sample particles to collection
chambers 600, where the particles are filtered out of the exhausted air,
i.e., separated from the exhausted air.

[0089] As most clearly illustrated in FIG. 9E, the seed and sample sorting
subsystem 570 still further includes a seed tray platform 602 adapted to
securely retain a plurality of the seed trays 18 in fixed positions and
orientations. Each seed tray 18 includes a plurality of seed wells 34,
each of which are adapted for receiving a seed after the respective seed
has been sampled by one of the grip and chip assemblies 512. For example,
in various embodiments, each seed tray 18 can be a twenty-four well tray.
The bank of grip and chip assemblies 512 is mounted to system support
structure above the seed tray platform 602 such that seeds can be
dispensed through the drain tubes 558 into selected seed wells 30 of
selected seed trays 18.

[0090] The seed tray platform 602 is mounted to an X-Y stage 604. The X-Y
stage 604 is a two-dimensional translation mechanism, including an X axis
translating track 606 and a Y axis translating track 608. The X-Y stage
604 additionally includes a first linear actuator 610 operable to
bidirectionally move a first carriage (not shown) along the length of the
X axis translating track 606. The X-Y stage 604 further includes a second
linear actuator 612 operable to bidirectionally move a second carriage
(not shown) along the length of the Y axis translating track 608. The Y
axis translating track 608 is mounted to the first carriage and the seed
tray platform 602 is mounted to the second carriage.

[0091] The first and second linear actuators 610 and 612 are controlled by
the CCS 700 to precisely move the seed tray platform 602 in two
dimensions. More particularly, the first and second actuators 610 and 612
move the seed tray platform 602 within an X-Y coordinate system to
precisely position any selected well 34 of any selected seed tray 18 at a
target location beneath a selected one or more of the drain tubes 558. In
various embodiments, the drain tubes 558 are held in linear alignment by
system support structure and the CCS 700 controls the first and second
actuators 610 and 612 to position a selected group of linearly adjacent
wells 34 at a target location beneath the linearly aligned drain tubes
558. More specifically, the CCS 700 moves the seed tray platform 602
within the X-Y coordinate system to position each of a plurality of
linearly adjacent wells 34 beneath a corresponding one of the linearly
aligned drain tubes 558. Therefore, when each of the seed gripping
mechanisms 522 releases the respective sampled seeds, each sampled seed
will fall, via gravity, vacuum or forced air, through the respective
drain tube 558 into the corresponding well 34 located beneath the
respective drain tube 558.

[0092] In operation, just prior to, simultaneously with, or just after the
set of seeds is retained by the seed gripping mechanisms 522, as
described above, the CCS 700 positions the selected well 34, or selected
group of wells 34, at the target location. Each seed is then sampled and
the samples are deposited in the sample tray wells 30, as described
above. Each seed gripping mechanism 522 is commanded to release the
respective seeds allowing the seeds to fall into the respective seed dump
bowls 554. Each seed dump bowl 554 funnels the respective seeds through
the respective drain port 556 and into the respective drain tubes 558.
The drain tubes 558 then direct the respective seeds into the selected
wells 34 positioned below the drain tubes 558. In various embodiments,
the one or more of the seeds can be sequentially released and the seed
tray platform 602 sequentially repositioned to deposit the one or more
seeds into selected wells 34. In other embodiments, the seed tray
platform is manipulated to position a group of linearly adjacent wells
beneath the linearly aligned drain tubes, all the seeds are then
substantially simultaneously released and deposited into the respective
group of linearly adjacent wells.

[0093] As the seed trays 18 are placed on the seed tray platform 602, a
tray identification number, e.g., a bar code, for each seed tray 18 and
the location of each seed tray 18 on the seed tray platform 602 is
recorded. Additionally, as each seed is deposited in a well 34, an X-Y
location of the well on the seed tray platform 602 can be recorded. The
recorded tray and well positions on the seed tray platform 602 can then
be compared to the X-Y locations of each deposited seed, to map the
specific seed in each well 34 of each seed tray 18. In various
embodiments, the seed tray platform 602 is removably coupled to the X-Y
stage 604 such that one or more seed tray platforms 602 can be loaded
with the seed trays 18 offline, i.e., away from the seed sorter system
10, and conveniently coupled to and decoupled from the X-Y stage 604.

[0094] As described above, each of the seed trays 18 and the sample trays
14 include a plurality of wells 34 and 30, respectively. In various
embodiments, the number and arrangement of the wells 34 in the seed trays
18 corresponds to the number and arrangement of the wells 30 in the
sample trays 14. This facilitates a one-to-one correspondence between a
seed and its extracted sample.

[0095] As described above, the sampling systems and methods of this
disclosure protect germination viability of the seeds so as to be
non-destructive. Germination viability means that a predominant number of
sampled seeds (i.e., greater than 50% of all sampled seeds) remain viable
after sampling. In some particular embodiments, at least about 75% of
sampled seeds, and in some embodiments at least about 85% of sampled
seeds remain viable. It should be noted that lower rates of germination
viability may be tolerable under certain circumstances or for certain
applications, for example, as genotyping costs decrease with time because
a greater number of seeds could be sampled for the same genotype cost.

[0096] In yet other embodiments, germination viability is maintained for
at least about six months after sampling to ensure that the sampled seed
will be viable until it reaches the field for planting. In some
particular embodiments, the methods of the present disclosure further
comprise treating the sampled seeds to maintain germination viability.
Such treatment may generally include any means known in the art for
protecting a seed from environmental conditions while in storage or
transport. For example, in some embodiments, the sampled seeds may be
treated with a polymer and/or a fungicide to protect the sampled seed
while in storage or in transport to the field before planting.

[0097] In various embodiments, the samples of the present disclosure are
used in a high-throughput, non-destructive method for analyzing
individual seeds in a population of seeds. The method comprises removing
a sample from the seed while preserving the germination viability of the
seed; and screening the sample for the presence or absence of one or more
characteristics indicative of a genetic or chemical trait. The method may
further comprise selecting seeds from the population based on the results
of the screening; and cultivating plants from the selected seed.

[0099] DNA may be extracted from the sample using any DNA extraction
methods known to those of skill in the art which will provide sufficient
DNA yield, DNA quality, and PCR response. A non-limiting example of
suitable DNA-extraction methods is SDS-based extraction with
centrifugation. In addition, the extracted DNA may be amplified after
extraction using any amplification method known to those skilled in the
art. For example, one suitable amplification method is the GenomiPhi®
DNA amplification prep from Amersham Biosciences.

[0100] The extracted DNA is screened for the presence or absence of a
suitable genetic marker. A wide variety of genetic markers are available
and known to those of skill in the art. The DNA screening for the
presence or absence of the genetic marker can be used for the selection
of seeds in a breeding population. The screening may be used to select
for QTL, alleles, or genomic regions (haplotypes). The alleles, QTL, or
haplotypes to be selected for can be identified using newer techniques of
molecular biology with modifications of classical breeding strategies.

[0101] In other various embodiments, the seed is selected based on the
presence or absence of a genetic marker that is genetically linked with a
QTL. Examples of QTLs which are often of interest include but are not
limited to yield, lodging resistance, height, maturity, disease
resistance, pest resistance, resistance to nutrient deficiency, grain
composition, herbicide tolerance, fatty acid content, protein or
carbohydrate metabolism, increased oil content, increased nutritional
content, stress tolerance, organoleptic properties, morphological
characteristics, other agronomic traits, traits for industrial uses,
traits for improved consumer appeal, and a combination of traits as a
multiple trait index. Alternatively, the seed can be selected based on
the presence or absence of a marker that is genetically linked with a
haplotype associated with a QTL. Examples of such QTL may again include,
without limitation, yield, lodging resistance, height, maturity, disease
resistance, pest resistance, resistance to nutrient deficiency, grain
composition, herbicide tolerance, fatty acid content, protein or
carbohydrate metabolism, increased oil content, increased nutritional
content, stress tolerance, organoleptic properties, morphological
characteristics, other agronomic traits, traits for industrial uses,
traits for improved consumer appeal, and a combination of traits as a
multiple trait index.

[0102] Selection of a breeding population could be initiated as early as
the F2 breeding level, if homozygous inbred parents are used in the
initial breeding cross. An F1 generation could also be sampled and
advanced if one or more of the parents of the cross are heterozygous for
the alleles or markers of interest. The breeder may screen an F2
population to retrieve the marker genotype of every individual in the
population. Initial population sizes, limited only by the number of
available seeds for screening, can be adjusted to meet the desired
probability of successfully identifying the desired number of
individuals. See Sedcole, J. R. "Number of plants necessary to recover a
trait." Crop Sci. 17:667-68 (1977). Accordingly, the probability of
finding the desired genotype, the initial population size, and the
targeted resulting population size can be modified for various breeding
methodologies and inbreeding level of the sampled population.

[0103] The selected seeds may be bulked or kept separate depending on the
breeding methodology and target. For example, when a breeder is screening
an F2 population for disease resistance, all individuals with the
desired genotype may be bulked and planted in the breeding nursery.
Conversely, if multiple QTL with varying effects for a trait such as
grain yield are being selected from a given population, the breeder may
keep individual identity preserved, going to the field to differentiate
individuals with various combinations of the target QTL.

[0104] Several methods of preserving single seed identity can be used
while transferring seed from a sampling facility to the field. Methods
include, but are not limited to, transferring selected individuals to
seed tape, a cassette tray, or indexing tray, transplanting with peat
pots, and hand-planting from individual seed packets. Multiple cycles of
selection can be utilized depending on breeding targets and genetic
complexity.

[0105] The screening methods of the disclosure may further be used in a
breeding program for introgressing a trait into a plant. Such methods
comprise removing a sample comprising cells with DNA from seeds in a
population, screening the DNA extracted from each seed for the presence
or absence of at least one genetic marker, selecting seeds from the
population based upon the results of the DNA screening; cultivating a
fertile plant from the seed; and utilizing the fertile plant as either a
female parent or male parent in a cross with another plant.

[0106] Examples of genetic screening to select seeds for trait integration
include, without limitation, identification of high recurrent parent
allele frequencies, tracking of transgenes of interest or screening for
the absence of unwanted transgenes, selection of hybrid testing seed, and
zygosity testing.

[0107] The identification of high recurrent pair allele frequencies via
the screening methods of the present disclosure again allows for a
reduced number of rows per population and an increased number of
populations, or inbred lines, to be planted in a given field unit. Thus,
the screening methods of the present disclosure may also effectively
reduce the resources required to complete the conversion of inbred lines.

[0108] The methods of the present disclosure further provide quality
assurance (QA) and quality control by assuring that regulated or unwanted
transgenes are identified and discarded prior to planting.

[0109] The methods of the present disclosure may be further applied to
identify hybrid seed for transgene testing. For example, in a conversion
of an inbred line at the BCnF1 stage, a breeder could effectively
create a hybrid seed lot (barring gamete selection) that was 50%
hemizygous for the trait of interest and 50% homozygous for the lack of
the trait in order to generate hybrid seed for testing. The breeder could
then screen all F1 seeds produced in the test cross and identify and
select those seeds that were hemizygous. Such method is advantageous in
that inferences from the hybrid trials would represent commercial hybrid
genetics with regard to trait zygosity.

[0110] Other applications of the screening methods of this disclosure for
identifying and tracking traits of interest carry the same advantages
identified above with respect to required field and labor resources.
Generally, transgenic conversion programs are executed in multi-season
locations which carry a much higher land and management cost structure.
As such, the impact of either reducing the row needs per population or
increasing the number of populations within a given field unit are
significantly more dramatic on a cost basis versus temperate
applications.

[0111] Still further, the screening methods of this disclosure may be used
to improve the efficiency of the doubled haploid program through
selection of desired genotypes at the haploid stage and identification of
ploidy level to eliminate non-haploid seeds from being processed and
advancing to the field. Both applications again result in the reduction
of field resources per population and the capability to evaluate a larger
number of populations within a given field unit.

[0112] In various embodiments, the disclosure further provides an assay
for predicting embryo zygosity for a particular gene of interest (GOI).
The assay predicts embryo zygosity based on the ratio of the relative
copy numbers of a GOI and of an internal control (IC) gene per cell or
per genome. Generally, this assay uses an IC gene that is of known
zygosity, e.g., homozygous at the locus (two IC copies per diploid cell),
for normalizing measurement of the GOI. The ratio of the relative copy
numbers of the IC to the GOI predicts the GOI copy number in the cell. In
a homozygous cell, for any given gene (or unique genetic sequence), the
gene copy number is equal to the cell's ploidy level since the sequence
is present at the same locus in all homologous chromosomes. When a cell
is heterozygous for a particular gene, the gene copy number will be lower
than the cell's ploidy level. The zygosity of a cell at any locus can
thus be determined by the gene copy number in the cell.

[0113] In some particular embodiments, the disclosure provides an assay
for predicting corn embryo zygosity. In corn seed, the endosperm tissue
is triploid, whereas the embryo tissue is diploid. Endosperm that is
homozygous for the IC will contain three IC copies. Endosperm GOI copy
number can range from 0 (homozygous negative) to 3 (homozygous positive);
and endosperm GOI copy number of 1 or 2 is found in seed heterozygous for
the GOI (or hemizygous for the GOI if the GOI is a transgene). Endosperm
copy number is reflective of the zygosity of the embryo: a homozygous
(positive or negative) endosperm accompanies a homozygous embryo,
heterozygous endosperm (whether a GOI copy number of 1 or 2) reflects a
heterozygous (GOI copy number of 1) embryo. The endosperm GOI copy number
(which can range from 0 to 3 copies) can be determined from the ratio of
endosperm IC copy number to endosperm GOI copy number (which can range
from 0/3 to 3/3, that is, from 0 to 1), which can then be used to predict
zygosity of the embryo.

[0114] Copy numbers of the GOI or of the IC can be determined by any
convenient assay technique for quantification of copy numbers, as is
known in the art. Examples of suitable assays include, but are not
limited to, Real Time (TaqMan®) PCR (Applied Biosystems, Foster City,
Calif.) and Invader® (Third Wave Technologies, Madison, Wis.) assays.
Preferably, such assays are developed in such a way that the
amplification efficiency of both the IC and GOI sequences are equal or
very similar. For example, in a Real Time TaqMan® PCR assay, the
signal from a single-copy GOI (the source cell is determined to be
heterozygous for the GOI) will be detected one amplification cycle later
than the signal from a two-copy IC, because the amount of the GOI is half
that of the IC. For the same heterozygous sample, an Invader® assay
would measure a GOI/IC ratio of about 1:2 or 0.5. For a sample that is
homozygous for both the GOI and the IC, the GOI signal would be detected
at the same time as the IC signal (TaqMan®), and the Invader assay
would measure a GOI/IC ratio of about 2:2 or 1.

[0115] These guidelines apply to any polyploid cell, or to haploid cells
(such as pollen cells), since the copy number of the GOI or of the IC
remain proportional to the genome copy number (or ploidy level) of the
cell. Thus, these zygosity assays can be performed on triploid tissues
such as corn endosperm.

[0116] The description herein is merely exemplary in nature and, thus,
variations that do not depart from the gist of that which is described
are intended to be within the scope of the teachings. Such variations are
not to be regarded as a departure from the spirit and scope of the
teachings.

Patent applications by Allen N. Ondes, St. Louis, MO US

Patent applications by Andrew M. Singleton, Manchester, MO US

Patent applications by Angela R. Koestel, St. Louis, MO US

Patent applications by Brian J. Forinash, St. Louis, MO US

Patent applications by Bruce J. Schnicker, Wildwood, MO US

Patent applications by David W. Finley, St. Louis, MO US

Patent applications by Heather M. Forbes, St. Charles, MO US

Patent applications by Jason K. Bull, Wildwood, MO US

Patent applications by Kevin L. Deppermann, St. Charles, MO US

Patent applications by Lee Yannakakis, Chesterfield, MO US

Patent applications by Sam R. Eathington, St. Louis, MO US

Patent applications by Monsanto Technology LLC

Patent applications in class CONDITION RESPONSIVE CONTROL PROCESS

Patent applications in all subclasses CONDITION RESPONSIVE CONTROL PROCESS